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Atdc5

Manufactured by RIKEN BioResource Center
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The ATDC5 is a cell line derived from mouse adipose tissue. It is capable of differentiating into adipocytes under appropriate culture conditions. The ATDC5 cell line can be used for the study of adipogenesis and related cellular processes.

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Lab products found in correlation

Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions) Dmem f12, by Thermo Fisher Scientific (2 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions) Pcdna3.1 vector, by Thermo Fisher Scientific (1 mentions) Atdc5, by Thermo Fisher Scientific (1 mentions) Ov nc, by Thermo Fisher Scientific (1 mentions) Lipofectamine 2000 transfection reagent, by Thermo Fisher Scientific (1 mentions) C3h10t1 2, by RIKEN BioResource Center (1 mentions) Penicillin streptomycin, by Nacalai Tesque (1 mentions) Streptomycin sulfate, by Merck Group (1 mentions) Penicillin, by Thermo Fisher Scientific (1 mentions) Streptomycin sulfate, by Thermo Fisher Scientific (1 mentions) Collagenase type 2, by Merck Group (1 mentions) Penicillin, by Merck Group (1 mentions) Ascorbate 2 phosphate, by Fujifilm (1 mentions)

Close spelling variants of this product

ATDC5 cell line (3 citations) ATDC5 cells (2 citations)

6 protocols using atdc5

1

Modulating Chondrocyte Inflammation with KT

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The chondrogenic cell line, ATDC5, was purchased from RIKEN BioResource Center and cultured in DMEM (Gibco; Thermo Fisher Scientific, Inc.) containing 10% FBS (Gibco; Thermo Fisher Scientific, Inc.) at 37˚C in a humidified atmosphere with 5% CO2. The cells were stimulated with IL-1β (10 ng/ml) for 24 h and then 5, 10 and 20 mg/ml KT were used to treat these IL-1β-induced ATDC5 cells at 37˚C for 24 h; IL-1β-induced ATDC5 cells without KT as used as the control.
With the aim of overexpressing COX-2 expression in ATDC5 cells, the pcDNA3.1 vector containing full-length COX-2 (OV-COX-2) and empty vector (OV-NC) were all designed and synthesized by Thermo Fisher Scientific, Inc. In addition, the cells that were not transfected with the plasmid were used as the control group. The transfection of ATDC5 cells was performed with Lipofectamine® 2000 transfection reagent (Invitrogen; Thermo Fisher Scientific, Inc.) at a concentration of 50 ng/ml. Following transfection for 48 h at 37˚C, the transfection efficiency was detected by reverse transcription-quantitative PCR (RT-qPCR) 48 h post-transfection.
Liu C, & Chen Y. (2022). Ketorolac tromethamine alleviates IL-1β-induced chondrocyte injury by inhibiting COX-2 expression. Experimental and Therapeutic Medicine, 23(5), 337.
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2

Isolation and Culture of Articular Chondrocytes

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We obtained samples of human articular cartilage from three individuals undergoing total knee arthroplasty after obtaining written informed consent from the patients. Human articular chondrocytes were isolated and cultured as previously described33 (link). C3H10T1/2, HuH-7and ATDC5 cells (Riken BRC, Tokyo, Japan) were maintained in monolayer culture as previously described34 (link). Primary mouse costal chondrocytes were isolated and the pellet cell cultures were performed. In addition, primary mouse articular chondrocytes were isolated and cultured as previously described35 (link) for use in functional analyses. Adenovirus vectors for GFP, Runx1, Bapx1 and Cre were prepared as previously described12 (link). Cells were transduced with adenoviral vectors at a multiplicity of infection (MOI) of 100. Lentivirus vectors of control shRNA (pSMART Non-targeting mCMV-TurboRFP) and Bapx1 shRNA (pSMART 2.0 mCMV/turboRFP Nkx3.2) were purchased from Dharmacon (Lafayette, CO).
Yano F., Ohba S., Murahashi Y., Tanaka S., Saito T, & Chung U.I. (2019). Runx1 contributes to articular cartilage maintenance by enhancement of cartilage matrix production and suppression of hypertrophic differentiation. Scientific Reports, 9, 7666.
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3

Chondrogenic Differentiation of ATDC5 Cells

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Since the procedure of obtaining normal cartilage from patients were not approved by the Medical Ethics Committee of Sun Yat-sen Memorial Hospital, we opted to use a chondrogenic cell line (ATDC5) purchased from Riken BioResource Center (Tsukuba, Japan). The cells were induced into normal chondrocytes by the addition of insulin, transferrin, and selenous acid (ITS, Sigma-Aldrich) according to previous studies17 (link).
Chen Z., Lin C.X., Song B., Li C.C., Qiu J.X., Li S.X., Lin S.P., Luo W.Q., Fu Y., Fang G.B., Wei-Ping L., Saw P.E, & Ding Y. (2020). Spermidine activates RIP1 deubiquitination to inhibit TNF-α-induced NF-κB/p65 signaling pathway in osteoarthritis. Cell Death & Disease, 11(7), 503.
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4

ATDC5 Chondrogenic Spheroid Formation

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Mouse chondrogenic progenitor cell line ATDC5 (RIKEN Bioresource Center, Japan) was cultured in DMEM/F-12 (Gibco, USA) containing 5% fetal bovine serum (Gibco, USA) and 1% penicillin-streptomycin (Nacalai Tesque, Japan) in a humidified incubator (37 , 5% CO 2 ). Cell passage was conducted every 3-4 days before reaching 80% -90% confluence. Spheroids were prepared by subculturing in a U-bottom ultra-low attachment 96-well plate (Thermo Fisher Scientific, USA) following previously reported methods (Kim et al. 2020 (Kim et al. , 2022)) . 2,500 cells were seeded to each well and incubated for 2, 4, 7, and 14 days, with medium changes every 2 -3 days.
Tomida K., Kim J., Maeda E., Adachi T, & Matsumoto T. (2024). Spatiotemporal analysis of multi-scale cell structure in spheroid culture reveals hypertrophic chondrocyte differentiation. Cell and tissue research.
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5

Culturing Chondrogenic Cells and Primary Chondrocytes

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The chondrogenic cell line ATDC5 was supplied by Riken Bio Resource Center (Tsukuba, Japan) and maintained in Dulbecco's modified Eagle's medium (DMEM)/F12 (Gibco, Carlsbad, CA, USA) with 5% fetal bovine serum (Gibco), 100 U/mL penicillin, and 100 mg/mL streptomycin sulfate (Life Technologies, Carlsbad, CA, USA) at 37 C under 5% CO 2 . Primary chondrocytes were harvested from the rib and articular cartilages dissected from newborn mice. After trypsin digestion, the primary chondrocytes were purified and digested with 0.1% collagenase type II (Sigma-Aldrich, St. Louis, MO, USA) with 10% FBS, 100 U/mL penicillin, and 100 mg/mL streptomycin sulfate at 37 C for 4 to 6 hours. The primary chondrocytes were resuspended, seeded into a 24-well plate, and cultured at 37 C under 5% CO 2 .
Li K., Yang P., Zhang Y., Zhang Y., Cao H., Liu P., Huang B., Xu S., Lai P., Lei G., Liu J., Tang Y., Bai X, & Zou Z. (2021). DEPTOR Prevents Osteoarthritis Development Via Interplay With TRC8 to Reduce Endoplasmic Reticulum Stress in Chondrocytes. Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research, 36(2).
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6

Murine ATDC5 Chondrogenic Differentiation Protocol

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The murine chondrogenic line ATDC5 was purchased from the Riken BioResource Center (Ibaraki, Japan). Cells were cultured in Dulbecco's Modified Eagle's Medium/Nutrient F-12 Ham (DMEM/ F12) (SigmaeAldrich, St Louis, MO) containing 5% FCS, 1% penicillin/ streptomycin (Thermo Fisher Scientific, Waltham, MA), 10 mg/ml human transferrin (SigmaeAldrich), 3 Â 10 À8 M sodium selenite (SigmaeAldrich), and 37.5 mg/ml ascorbate 2-phosphate (Wako, Osaka, Japan), as described 16e18 . Cells were maintained at 37 C in a humidified atmosphere of 5% CO 2 in air for the entire culture period. To induce chondrogenic differentiation, 10 mg/ml human insulin (SigmaeAldrich) was added to the medium and supplemented to cultures when medium was changed.
Tanoue H., Morinaga J., Yoshizawa T., Yugami M., Itoh H., Nakamura T., Uehara Y., Masuda T., Odagiri H., Sugizaki T., Kadomatsu T., Miyata K., Endo M., Terada K., Ochi H., Takeda S., Yamagata K., Fukuda T., Mizuta H, & Oike Y. (2018). Angiopoietin-like protein 2 promotes chondrogenic differentiation during bone growth as a cartilage matrix factor. Osteoarthritis and cartilage, 26(1).
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